Noritake M, Katsura Y, Shinomiya N, Kanatani M, Uwabe Y, Nagata N, Tsuru S
Third Department of Internal Medicine, National Defence Medical College, Saitama, Japan.
Clin Exp Immunol. 1992 May;88(2):269-74. doi: 10.1111/j.1365-2249.1992.tb03072.x.
Although the standard assays for reactive oxygen species have been based on the measurement of those released into the extracellular environment, the microbicidal capacity to the engulfed microorganisms is mainly dependent on those released into the intracellular environment, such as phagosomes. We studied intracellular oxidative activities of individual phagocytes by dichlorofluorescein (DCFH) oxidation assay to investigate the relationship between the reactive oxygen species released intracellularly and the impaired microbicidal capacity in diabetic patients. Time courses of intracellular production of hydrogen peroxide by polymorphonuclear leucocytes (PMNL) and monocytes were observed at the resting condition and after the stimulation with phorbol myristate acetate (PMA; 160 nM) by flow cytometry. Thirty-four patients with non-insulin-dependent diabetes mellitus (NIDDM) and 23 age-matched healthy volunteers were subjected to the studies. PMNL from patients with NIDDM showed a significantly decreased capacity to produce hydrogen peroxide after the stimulation (P less than 0.05 at 15 min, P less than 0.01 at 30 and 45 min). By contrast, intracellular hydrogen peroxide production by monocytes at the resting condition and an early stimulatory phase (8 min after the stimulation) was significantly (P less than 0.01) enhanced in patients with NIDDM compared with that in controls. Both the changes of intracellular hydrogen peroxide production observed in PMNL and monocytes from patients with NIDDM were in association with an increased haemoglobin Alc level in erythrocytes, but did not relate to total cholesterol and triglyceride levels in the serum. The possible mechanisms of these dissociated changes in hydrogen peroxide producing capacity of phagocytes from patients with NIDDM are discussed.
尽管活性氧的标准检测方法一直基于对释放到细胞外环境中的活性氧的测量，但对被吞噬微生物的杀菌能力主要取决于释放到细胞内环境中的活性氧，如吞噬体。我们通过二氯荧光素（DCFH）氧化试验研究了单个吞噬细胞的细胞内氧化活性，以探讨细胞内释放的活性氧与糖尿病患者杀菌能力受损之间的关系。通过流式细胞术观察了多形核白细胞（PMNL）和单核细胞在静息状态下以及在用佛波酯肉豆蔻酸酯（PMA；160 nM）刺激后的细胞内过氧化氢产生的时间进程。34例非胰岛素依赖型糖尿病（NIDDM）患者和23例年龄匹配的健康志愿者参与了研究。NIDDM患者的PMNL在刺激后产生过氧化氢的能力显著降低（15分钟时P<0.05，30和45分钟时P<0.01）。相比之下，与对照组相比，NIDDM患者单核细胞在静息状态和早期刺激阶段（刺激后8分钟）的细胞内过氧化氢产生显著增强（P<0.01）。在NIDDM患者的PMNL和单核细胞中观察到的细胞内过氧化氢产生的变化均与红细胞中糖化血红蛋白Alc水平的升高有关，但与血清中的总胆固醇和甘油三酯水平无关。本文讨论了NIDDM患者吞噬细胞过氧化氢产生能力这些解离变化的可能机制。
